Improved Loading of Plasma-Derived Extracellular Vesicles to Encapsulate Antitumor miRNAs
Open accessAbstract
Extracellular vesicles (EVs) carry various molecules involved in intercellular communication and have raised great interest as drug delivery systems. Several engineering methods have been investigated for vesicle loading. Here, we studied the electroporation of EVs isolated from plasma to load antitumor microRNAs (miRNAs). First, we optimized the transfection protocol using miRNA cel-39 by evaluating different parameters (voltage and pulse) for their effect on vesicle morphology, loading capacity, and miRNA transfer to target cells. When compared with direct incubation of EVs with miRNA, mild electroporation allowed more efficient loading and better protection of miRNA from RNase degradation. Moreover, electroporation preserved the naive vesicle cargo, including RNAs and proteins, and their ability to be taken up by target cells, supporting the absence of vesicle damage. EVs engineered with antitumor miRNAs (miR-31 and miR-451a) successfully promoted apoptosis of the HepG2 hepatocellular carcinoma cell line, silencing target genes involved in anti-apoptotic pathways. Our findings indicate an efficient and functional miRNA encapsulation in plasma-derived EVs following an electroporation protocol that preserves EV integrity.
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Electroporation has been the primary method utilized for loading RNAs into exosomes.
“Electroporation has been the primary method utilized for loading RNAs into exosomes.”
From Exosome for mRNA delivery: Strategies and therapeutic applications (2024) · cites this paper as [107]
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- Authors
- Pomatto, M. A. C., Bussolati, B., D’Antico, S., Ghiotto, S., Tetta, C., Brizzi, M. F., & Camussi, G.
- Affiliations
- University of Turin (IT)
- Azienda Ospedaliera Citta' della Salute e della Scienza di Torino (IT)
- Corresponding author
- Camussi, Giovanni
- Venue
- Molecular Therapy - Methods & Clinical Development
- Publisher
- Elsevier BV
- Published
- 2019-06
- Volume
- 13
- Pages
- 133-144
- ISSN
- 2329-0501 (ISSN-L)
- PMC
- PMC6370572
- OpenAlex
- W2910555711
- Cited by
- 243
- References
- 62
- Citations by year
- 2026: 27 · 2025: 35 · 2024: 34 · 2023: 33 · 2022: 29 · 2021: 44 · 2020: 31 · 2019: 10
- FWCI
- 9.43
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Molecular Therapy — Methods & Clinical Development (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Institutional Research Information System University of Turin (University of Turin) (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- Associazione Italiana per la Ricerca sul Cancro (IG2012)
- Regione Piemonte (POR FESR 2014/2020)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Keywords
- extracellular vesicles, plasma-derived vesicles, electroporation, miR-31, miR-451a, vesicle loading, RNase protection, HepG2 cells, hepatocellular carcinoma, apoptosis, drug delivery systems, vesicle integrity
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